Chen Yang-Yi, Huang Shu-Mei, Cheng Yu-Wen, Yen Meng-Chi, Hsu Ya-Ling, Lan Cheng-Che E
Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.
Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
J Dermatol Sci. 2023 Jan;109(1):37-46. doi: 10.1016/j.jdermsci.2023.01.007. Epub 2023 Jan 18.
Impaired wound healing is a serious diabetes complication compromising patients' quality of life. However, the pathogenesis of diabetic wounds (DWs) remains incompletely understood. Human epidermal keratinocyte (HEK) is the sentinel cell that initiates healing processes after the epidermal integrity has been disrupted.
This study aimed to investigate the functional roles of HEKs in wound healing and to identify candidate genes, signaling pathways and molecular signatures contributing to the DWs.
HEKs were cultured in normal or high-glucose environment, followed by scratch, to mimic the microenvironment of normal wounds and DWs. Subsequently, we performed RNA sequencing and systematically analyzed the expression profiles by bioinformatics approaches.
High-glucose environment altered the keratinocyte transcriptome responses to wounding. In experimental model of DWs, we found that TNF, CYP24A1, NR4A3 and GGT1 were key overexpressed genes in keratinocytes and were implicated in multiple cellular responses. Further analysis showed that wounding in high-glucose environment activated G-protein-coupled receptor (GPCR) signaling, cAMP response element-binding protein (CREB) signaling, and adrenomedullin signaling in keratinocytes, while dysregulated skin development and immune responses as compared to their counterpart in normal glucose settings.
This simplified in-vitro model serves as a valuable tool to gain insights into the molecular basis of DWs and to facilitate establishment of personalized therapies in clinical practice.
伤口愈合受损是一种严重的糖尿病并发症，会影响患者的生活质量。然而，糖尿病伤口（DWs）的发病机制仍未完全明确。人类表皮角质形成细胞（HEK）是在表皮完整性被破坏后启动愈合过程的哨兵细胞。
本研究旨在探讨HEKs在伤口愈合中的功能作用，并确定导致DWs的候选基因、信号通路和分子特征。
将HEKs在正常或高糖环境中培养，然后进行划痕处理，以模拟正常伤口和DWs的微环境。随后，我们进行了RNA测序，并通过生物信息学方法系统地分析了表达谱。
高糖环境改变了角质形成细胞对伤口的转录组反应。在DWs实验模型中，我们发现TNF、CYP24A1、NR4A3和GGT1是角质形成细胞中关键的过表达基因，并参与多种细胞反应。进一步分析表明，与正常葡萄糖环境中的对应物相比，高糖环境中的伤口激活了角质形成细胞中的G蛋白偶联受体（GPCR）信号、环磷酸腺苷反应元件结合蛋白（CREB）信号和肾上腺髓质素信号，同时皮肤发育和免疫反应失调。
这种简化的体外模型是深入了解DWs分子基础以及促进临床实践中个性化治疗方案制定的宝贵工具。
